alexa Analysis of recombinant monoclonal antibodies by capillary zone electrophoresis.
Chemical Engineering

Chemical Engineering

Journal of Chromatography & Separation Techniques

Author(s): Espinosade la Garza CE, PerdomoAbndez FC, PadillaCaldern J, UribeWiechers JM, Prez NO,

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Abstract Analytical platforms that characterize charge heterogeneity in therapeutic proteins, such as mAbs, are important tools that can be used to define quality attributes. CZE separates protein moieties close to their native state and is a valuable physicochemical analytical method that can be used in parallel with other orthogonal methods for characterization and comparability. In this study, custom conditions for the analysis of charge heterogeneity of two mAbs were developed with regard to critical parameters in the BGE, running conditions, and sample treatment. The method application was tested for up to four mAbs and one mAb fragment. The electropherograms showed specific profiles and contrasting levels of basic and acidic isoforms with respect to the main isoform. Issues that surround this method, such as peak tailing and capillary lifetime, are summarized. Using this method, the identities of rituximab and trastuzumab were confirmed, based on the correspondence between the biosimilars and reference products, noninterference of the sample matrix, and the ability to separate spiked samples of related mAbs. The RSD of the isoform content and migration time for the method repeatability were less than 2 and 1\%, respectively. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. This article was published in Electrophoresis and referenced in Journal of Chromatography & Separation Techniques

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